5beta-cyano-a-norandrostane compounds



3,346,616 SB-CYANO-A-NORANDROSTANE COMPOUNDS Seymour D. Levine, North Brunswick, and Pacifico A. Principe, South River, N.J., assignors to E. R. Squibb & Sons, Inc., New York, N.Y., a corporation of Delaware No Drawing. Filed Aug. 29, 1966, Ser. No. 575,515 9 Claims. (Cl. 260-464) This invention relates to new steroidal compounds and, more particularly, to new hydroxylated and acylated steroids of the A-norandrostane series, new intermediates useful in the preparation of the same, and processes for preparing the same.

The new final products of this invention are of the Formula 1:

wherein R and R are hydroxy or acyloxy and R is selected from the group hydrogen, vinyl, ethynyl, halo substituted ethynyl and trifluoromethyl substituted ethynyl, and R and R together are x0 (0 Among the suitable acyloxys may be mentioned the acyloxy radical of a hydrocarbon carboxylic acid of less than twelve carbon atoms, as exemplified by, the lower alkanoic acids (e.g., acetic, propionic, butyric, hexanoic and en-anthic acid), the lower alkenoic acids, the. cycloalkane carboxylic acids, the cycloalkene carboxylic acids, the monocyclic aromatic carboxylic acids (e.g., benzoic acid) and the monocyclic aryl-(lower alkanoic) acids (e.g., phenacetic and ,B-phenylpropionic acid). Among the suitable halogen substituted vinyls may be mentioned perhalovinyls, such as trifluorovinyl, trichlorovinyl, 1,2-difiuoro-2-chlorovinyl and 1,2-difluoro 2 bromovinyl; the dihalovinyls, such as 1,2-difluorovinyl, 2,2-difluorovinyl, 1-chl0ro-2-flu0rovinyl, l-bromo-2-fiuorovinyl, and 1,2-dichlorovinyl; and the monohalovinyls, such as l-fluorovinyl, Z-fiuorovinyl, l-chlorovinyl, 2-chlorovinyl, and .lbromovinyl. Among the suitable halogen substituted ethynyls may be mentioned fiuor-oethynyl, chloroethynyl and bromoethynyl.

, The final products of this invention are physiologically active compounds that possess anti-androgenic activity (i.e., they can be utilized in the treatment of such conditions as hyperandrogenic acne, the dose being adjusted for the activity of the particular compound.

The compounds of this invention are prepared by reducing a compound having the Formula II:

CN n

where R and R are as hereinbefore defined. The 2- hydroxy-17-acyloxy and 2,17-diol compounds of this invention are prepared by treating the corresponding 2-oxo starting material with a reducing agent such as an alkali United States. Patent 0 metal borohydride, e.g., sodium borohydride, potassium borohydride or lithium borohydride to form a mixture of the 2u-hydroxy and ZB-hydroxy derivatives which are then separated by chromatography. The 17-oxo compounds of this invention may be prepared by subjecting 5,8-cyano-A-norandrostane 2 one-17B-ol to the action of the microorganism Fusarium javanz'cum. This treatment surprisingly yields the 17.-oxo 2a hydroxy compounds of the invention. The conditions for such microbial reaction are well known in the art and are similar to those specified in US. Patent No. 3,179,698.

Starting materials of this invention are prepared by interacting a compound of the Formula III:

RI, CN III wherein 'R" is ketal, with a compound of the formula: R X, wherein R is as hereinbefore defined, and X is an activating group such as magnesium bromide, lithium or a complex thereof (e.g., a lithium ethylenediamine complex). Although any ketal may beused, the preferred ketals are those with alkanediols, such as ethylene glycol and propylene glycol. To complete the reaction, water is then added, thereby yielding products of the Formula 1V:

RII CN 7 wherein R and R are as hereinbefore defined. The compound of Formula 1V formed contains a 2- ketal group. It may then be hydrolyzed, by treatment with p-toluenesulfonic acid, in acetone-water to yield the corresponding 2-keto derivative starting material of the instant invention. 1

Preparation of the starting material-s of the instant invention wherein R is hydroxy and R is hydrogen may be prepared in copending application, Ser. No. 355,913, filed Mar. 30, 1964, now U.S. Patent, No. 3,271,437. If a 17-ester is desired as the final product, the compound of Formula IV, in its free Z-keto form, is reacted with the acyl chloride or acid anhydride of the desired acid, preferably one of the acids mentioned hereinbefore, to yield the l7-ester.

The following examples illustrate temperatures being in centigrade):

EXAMPLE 1 Za-hydrOxy-Sfl-cyzunO-A-norandr0stane-17-one the invention (all (A) Fermentation.Surface growth from each of 2 two week old agar slants of Fusarium javanicum var. ensiforme (QM-524) (Army Quartermaster, Natick, Mass.),

the slants containing as a nutrient medium (A):

Grams K HPO 1 Agar 20 Distilled water to 1 liter is suspended in 5 ml. of 0.01% aqueous sodium lauryl sulfate solution. One ml. portions of this suspension are used to inoculate eight 250 ml. Erlenmeyer flasks each containing 50 ml. of the following sterilized medium (B):

Distilled water to 1 liter.

After seventy-two hours incubation at 25 C. with continuous rotary agitation (280 cycles/minute; two-inch stroke), 10% (vol./vol.) transfers are made to thirtyfour 250 ml. Erlenmeyer flasks each containing 50 ml. of freshly sterilized medium B. After twenty-four hours of further incubation, using the same conditions as described above the steroid (300 micrograms/ml.) is added by supplementing each flask with 0.25 ml. of a sterile solution (60 mg./ml.) of 5/3-cyano-A-norandrostane-2-one-17fi-ol in N,N-dirnethylformamide. A total of 510 mg. is fermented.

After seventy-two hours of further incubation, using identical conditions as described above, the contents of the flasks are pooled and the broth is then filtered through a Seitz clarifying pad. The flasks, mycelium and pad are washed with successive 50 ml. portions of warm water. The combined filtrate and washings have a volume of 2000 m1.

(B) lslati0n.The thus obtained filtrate is extracted three times with chloroform. The chloroform extracts are Washed three times with water, dried over sodium sulfate and evaporated to dryness. The residue is crystallized from methanol-isopropyl ether to give 242 mg. of 2a hydroxy /8-cyano-A-norandrostane-17-one having a melting point of 260-261. The analytical sample is recrystallized from chloroform-isopropyl ether, M.P. 264- 265"; [a] +93 (EtOH); A 2.92, 4.51 and 5.78;;

1%3 ,9.13 (s., 18-Me) 8.74 (s., 19-Me) and 5.65 (m., 25-H).

Analysisr Calcd for C19H2'702N (301.41): C, 75.71; H, 9.03; N, 4.65. Found: C, 75.70; H, 9.16; N, 4.42. EXAMPLE 2 2ot-acet0xy-5/3-cyan0-A -norandr0stane-1 7 -one A solution of 50 mg. of 2u-hydroxy-5fl-cyano-A-norandrostane-17-one in 0.4 ml. of acetic anhydride and 0.8 ml. of pyridine is warmed on a steam bath for 1.25 hours, diluted with Water and extracted with chloroform. The chloroform extracts are washed with 8% salt solution, dried over sodium sulfate, and evaporated. The

residue is crystallized from ether-isopropyl ether to give 30mg. of 2a-acetoxy-5,8-cyano-A-norandrostane-17-one, M.P. 159160. The analytical sample is prepared by recrystallization from chloroform-isopropyl ether, M.P. 159160; A 4.48 and 5.77 75%,, 9.13 (s., 18-Me) 8.73 (s., 19-Me), 7.95 (s., 2a-acetate) and 4.87 (m., J =24 cps., 2 8-H).

Analysis.-Calcd for C H O N (343.45): C, 73.43; H, 8.51; N, 4.08. Found: C, 72.62; H, 8.49; N, 4.20. EXAMPLE 3 Za-hydroxy-S/i-cyano-l 7,3-acet0xy-A -n0randrostane and 2 fl-hydroxy-S 13-cyan0-1 7 fl-acetoxy-A -n0randr0stane A solution of 444 mg. of 5,6-cyano-17fi-acetoxy-A-norandrostane-Z-one in 20 ml. of methanol is treated with 200 mg. of sodium borohydride and left at room temperature for two hours. The reaction mixture is treated with 2 N HCl and water and the methanol removed in vacuo. The aqueous phase is extracted with chloroform and the chloroform extracts washed with 8% salt solution, dried over sodium sulfate and evaporated to dryness. Plate chromatography of the residue using neutral alumina (Activity V) as the adsorbent and chloroform 2% methanol as the developing solvent gives a diffuse band which is detectable by iodine. The band is arbitrarily divided in two portions. Elution of the more polar portion with ethyl acetate and crystallization of the residue from chloroform-ether gives 136 mg. of ZtX-hy'dI'OXY- Sfl-cyano-17B-acetoxy-A-norandrostane having a melting point of 246-248. The analytical sample is prepared by recrystallization from chloroform-isopropyl ether, 24-6.5 247.5"; A 2.87, 4.48 and 5.78

5 9.20 (s., 18-Me) 8.75 (s., l9-Me), 7.97 (s., H S-acetate), 5.55 (m., ZB-H) and 5.42 (m., l7a-H).

Analysis.Calcd for C H O N (345.47): C, 73.00; H, 9.05; N, 4.05. Found: C, 73.70; H, 8.98; N, 3.89.

The more polar portion is eluted with ethyl acetate and the residue rechromatographed on neutral alumina (Activity V) using chloroform as the developing solvent to give 2 major bands. The less polar band is eluted with ethyl acetate to give after evaporation and crystallization from chloroform-isopropyl ether 2,8-hydroxy-5fi-cyano- 17fi-acetoxy-A-norandrostane, M.P. 174-175 The analytical sample is prepared by recrystallization from chloroform-isopropyl ether, M.P. 175-176; A 2.87, 4.50 and 5.78

.gg g, 9.20 (s., 18-Me) 8.83 (s., l9-Me); 7.96 (s., l7/3-acetate), 5.42 (m., 17a-H and 2a-H).

Analysis.Calcd for C H O N (345.47): C, 73.00; H, 9.05; N, 4.05. Found: C, 73.01; H, 9.06; N, 4.25.

EXAMPLE 4 2a,] 7fl-diacetoxy-5p-cyano-A-n0randr0stane A solution of mg. of 2a-hydroxy-5fi-cyano-17[3 acetoxy-A-norandrostane in 1 ml. of acetic anhydride and 2 ml. of pyridine is left overnight at room temperature. The reaction mixture is diluted With water and the precipitate collected by filtration to give 103 mg. of 212,175- diacetoxy-Sfi-cyano-A-norandrostane having a melting point of 146-147.5. The analytical sample is prepared by recrystallization from isopropyl ether, M.P. 1495- 150; A 4.49, 5.80m;

75 3 3 9.22 (s., 18-Me) 8.75 (s., l9-Me), 7.98 (s., Zoe-acetate), 7.95 (s., acetate), 5.46 (t., 17oc-H) and 4.86 (J =26 cps., 2 8-H). Analysis.Calcd for C H O N (387.50): C, 71.29; H, 8.58; N, 3.61. Found: C, 71.03; H, 8.65; N, 3.68.

EXAMPLE 5 25,1 7 p-diacetOxy-S fi-cyano A -n0randr0stane Following the procedure in Example 4 but substituting 2/3 hydroxy SB-cyano-17fi-acetoxy-A-norandrostane for the Zea-isomer there is obtained 2,8,17,8-diacetoxy-5flcyano-A-norandrostane.

EXAMPLE 6 1 2a,17fi-dihydroxy-5fi-cyano-A-ri0randrostane 5 cyano-A-norandrostane, M.P. 215-217. The analytical sample is prepared by recrystallization from methanolisopropyl ether, M.P. 221-222"; A 2.93 and 4.50 Analysis.-Calcd for C H O N (303.43): C, 75.20; H, 9.63; N, 4.62. Found: C, 75.17; H, 9.37; N, 4.80.

EXAMPLE 7 Following the procedure of Example 3 but substituting 17u-ethynyl-Sfi-cyano-A-norandrostane 2 one-17fi-ol in lieu of SB-cyano-l7fi-acetoxy-A-norandrostane-Z-one the desired products are obtained.

EXAMPLE 8 1 7a-ethyny l-Sfl-cyano-A -nrandr0stane-2 a-hydr0xy 1 75-01 17-acetate Substituting 17oz ethynyl-S 3-cyano-A-norandrostane-Z- one-17/3-o1 acetate in lieu of SB-cyano-l7/3-acetoXy-A-norandrostane-Z-one in Example 3 the desired product is obtained.

EXAMPLE 9 Following the procedure of Example 3 but utilizing 17oz trifiuoropropynyl-fi-cyano-A-norandrostane-2-one- 17 8-01 in lieu of 5,8-cyano-17}?)-acetoXy-A-norandrostane- 2-one the desired products are obtained.

In a similar manner following the procedure of Example 3 but utilizing 17oc-ChlOIO6thY11Y1-5 B-cyano-A-norandrostane-2-one- 1fi ifiroethynyl-SBcyano-A-norandrostane-Z-one- 17itif l i1 orovinyl-5 fi-cyano-A-norandrostane-2-one- 17i EZlilOroVinyI-Sfl-cyam-A-norandrostane-2-one- 17 iziiichlorovinyldfi-cyano-A-norandrostane-Z-one- 17 iih ciifluorovinyl-5 fl-cyano-A-norandrostane-2-one- 17i t rig1 1 oropropynyl-5 ,8-cyano-A-norandrostane-2-one- 175-01 acetate;

17a-ethynyl-SB-A-norandrostane-Z-one-17,8-01

the products formed are:

17 a-chloroethynyl-SB-cyano-A-norandrostane-2fi, 1 7,8-diol and 17a-chloroethynyl-SB-cyano-A-norandrostane-2a,175-

diol;

17/3-ch1or0ethynyl-5 B-cyano-A-norandrostane-2 3, 17 ,8-

diol and 17 fi-chloroethynyl-S B-cyano-A-norandrostane-2 a, 17 pdiol;

17a-trifluorovinyl-Sfi-cyano-A-norandrostane-213,17,8-diol and 17a-trifiuorovinyl-5 3-cyano-A-norandrostane-2a, 1713- diol;

6 17ot-B-chlorovinyl-5B-cyano-A-norandrostane-2,B,17,8-

diol and 17a-fi-chlorovinyl-5fi-cyano-A-norandrostane-h, 17,6-

diol; 17(it-0 8-dichlorovinyl-5p-cyano-A-norandrostane2,8,17B-

diol and 17a-a, 8-dichlorovinyl-SB-cyano-A-norandrostane-2a,17B-

diol; 17a-a,fi-difiuorovinyl-5,B-cyano-A-norandrostane-ZB, 17 8- diol and 17a-et,,8-difluorovinyl-5,8-cyano-A-norandrostane-2a,17,8-

diol; 17a-trifluoropropynyl-SB-cyano-A-norandrostane-2,B, 17,8-

diol and l7a-trifluoropropynybSfl-cyano-A-norandrostane-2a,176-

diol; respectively.

The invention may be otherwise embodied within the scope of the appended claims.

We claim: 1. A compound having the formula References Cited UNITED STATES PATENTS 3,271,437 9/1966 Levine 260-464 CHARLES B. PARKER, Primary Examiner.

JOSEPH P. BRUST, Examiner, 

1. A COMPOUND HAVING THE FORMULA 